Photosynthetic Picoeukaryotes Research Articles

Growth and mortality rates of picophytoplankton in the Baltic Sea Proper

Picophytoplankton (<2 µm diameter), a diverse group of picocyanobacteria and photosynthetic picoeukaryotes, are significant contributors to primary production. Predatory mortality controls picophytoplankton biomass and thereby energy transfer in the marine food web. The 2 major pathways of picophytoplankton mortality are grazing and viral lysis. Grazing passes carbon directly to higher trophic levels, while lysis products are passed into the viral loop. Picophytoplankton are abundant in the Baltic Sea but little is known about their predatory mortality. Using a modification of the dilution approach, we calculated growth and mortality rates of picophytoplankton and studied the effect of predation on community structure during late August and September. The experiments were conducted coinciding with the peak in picophytoplankton abundance (∼105 cells ml-1) at the Linnaeus Microbial Observatory in the Baltic Sea Proper. The results showed that grazing is an important controller of picocyanobacteria and photosynthetic picoeukaryote populations, while no significant viral lysis effect was detected. Grazing on picocyanobacteria was proportional to growth rates, while grazing on photosynthetic picoeukaryotes exceeded growth. Selective grazing of phylogenetically distinct picocyanobacterial clades had a significant effect on community structure, suggesting that grazing has an impact on the seasonal dynamics of co-occurring clades. Picocyanobacteria had a higher carbon transfer contribution to higher trophic levels than photosynthetic picoeukaryotes at the time of the experiments. The study shows that picophytoplankton are important contributors to carbon cycling in the Baltic Sea microbial food web and should be considered for future ecological models.

Chromosome-scale assembly of the streamlined picoeukaryote Picochlorum sp. SENEW3 genome reveals Rabl-like chromatin structure and potential for C4 photosynthesis.

Genome sequencing and assembly of the photosynthetic picoeukaryotic Picochlorum sp. SENEW3 revealed a compact genome with a reduced gene set, few repetitive sequences, and an organized Rabl-like chromatin structure. Hi-C chromosome conformation capture revealed evidence of possible chromosomal translocations, as well as putative centromere locations. Maintenance of a relatively few selenoproteins, as compared to similarly sized marine picoprasinophytes Mamiellales, and broad halotolerance compared to others in Trebouxiophyceae, suggests evolutionary adaptation to variable salinity environments. Such adaptation may have driven size and genome minimization and have been enabled by the retention of a high number of membrane transporters. Identification of required pathway genes for both CAM and C4 photosynthetic carbon fixation, known to exist in the marine mamiellale pico-prasinophytes and seaweed Ulva, but few other chlorophyte species, further highlights the unique adaptations of this robust alga. This high-quality assembly provides a significant advance in the resources available for genomic investigations of this and other photosynthetic picoeukaryotes.

Micromonas, a small pigmented flagellate, predominates the nanoflagellate and photosynthetic picoeukaryote communities in the northern South China Sea.

A small pigmented flagellate, Micromonas, is prevalently distributed in coastal and pelagic waters. However, there have been few studies conducted to quantify their abundance in the marginal seas of the Northwest Pacific Ocean. In this study, we used fluorescent in situ hybridization with tyramide signal amplification (TSA-FISH) to reveal the spatial distribution of Micromonas in the northern South China Sea (SCS). On average, the abundance of Micromonas was 317 cells mL-1, with the average proportions in the nanoflagellates (NF) and photosynthetic picoeukaryotes (PPE) communities being 10.94% and 15.39%, respectively. This indicates a wide distribution and dominance of this genus in the studied area. The relationships between Micromonas abundance and various environmental factors suggested that biotic correlations play more important roles than physicochemical filtering on Micromonas assemblage. This may indicate a broad environmental adaptation spectrum of this genus through its flexibility in terms of resource acquisition strategies. In summary, this study provides insight into the spatial distribution pattern of Micromonas and highlights its crucial contribution to the composition of NFs and PPE communities, which rely on biological interaction to respond to the changing environmental conditions in the northern SCS.

Genomic architecture constrains macromolecular allocation in dinoflagellates

Dinoflagellate genomes have a unique architecture that may constrain their physiological and biochemical responsiveness to environmental stressors. Here we quantified how nitrogen (N) starvation influenced macromolecular allocation and C:N:P of three photosynthetic marine dinoflagellates, representing different taxonomic classes and genome sizes. Dinoflagellates respond to nitrogen starvation by decreasing cellular nitrogen, protein and RNA content, but unlike many other eukaryotic phytoplankton examined RNA:protein is invariant. Additionally, 2 of the 3 species exhibit increases in cellular phosphorus and very little change in cellular carbon with N-starvation. As a consequence, N starvation induces moderate increases in C:N, but extreme decreases in N:P and C:P, relative to diatoms. Dinoflagellate DNA content relative to total C, N and P is much higher than similar sized diatoms, but similar to very small photosynthetic picoeukaryotes such as Ostreococcus. In aggregate these results indicate the accumulation of phosphate stores may be an important strategy employed by dinoflagellates to meet P requirements associated with the maintenance and replication of their large genomes.

Genetic and physiological responses to light quality in a deep ocean ecotype of Ostreococcus, an ecologically important photosynthetic picoeukaryote.

Phytoplankton are exposed to dramatic variations in light quality when cells are carried by upwelling or downwelling currents or encounter sediment. We investigated the potential impact of light quality changes in Ostreococcus, a key marine photosynthetic picoeukaryote, by analysing changes in its transcriptome, pigment content, and photophysiology after acclimation to monochromatic red, green, or blue light. The clade B species RCC809, isolated from the deep euphotic zone of the tropical Atlantic Ocean, responded to blue light by accelerating cell division at the expense of storage reserves and by increasing the relative level of blue-light-absorbing pigments. It responded to red and green light by increasing its potential for photoprotection. In contrast, the clade A species OTTH0595, which originated from a shallow water environment, showed no difference in photosynthetic properties and minor differences in carotenoid contents between light qualities. This was associated with the loss of candidate light-quality responsive promoter motifs identified in RCC809 genes. These results demonstrate that light quality can have a major influence on the physiology of eukaryotic phytoplankton and suggest that different light quality environments can drive selection for diverse patterns of responsiveness and environmental niche partitioning.

Phytoplankton Diversity and Co-Dependency in a Stratified Oligotrophic Ecosystem in the South Adriatic Sea

The oligotrophy of the southern Adriatic Sea is characterized by seasonal stratification which enables nutrient supply to the euphotic layer. A set of interdisciplinary methods was used to elucidate the diversity and co-dependency of bacterio- and phytoplankton of the water column during the stratification period of July 2021. A total of 95 taxa were determined by microscopy: 58 diatoms, 27 dinoflagellates, 6 coccolithophores, and 4 other autotrophs, which included Chlorophyceae, Chrysophyceae, and Cryptophytes. Nanophytoplankton abundances were higher in comparison to microphytoplankton. The prokaryotic plankton community as revealed by HTS was dominated by Proteobacteria (41–73%), Bacteroidota (9.5–27%), and cyanobacteria (1–10%), while the eukaryotic plankton community was composed of parasitic Syndiniales (45–80%), Ochrophyta (2–18%), Ciliophora (2–21%), Chlorophytes (2–4%), Haptophytes (1–4%), Bacillariophyta (1–13%), Pelagophyta (0.5–12%) and Chrysophyta (0.5–3%). Flow cytometry analysis has recorded Prochlorococcus and photosynthetic picoeukaryotes as more abundant in deep chlorophyll maximum (DCM), and Synechococcus and heterotrophic bacteria as most abundant in surface and thermocline layers. Surface, thermocline, and DCM layers were distinct considering community diversity, temperature, and nutrient correlations, while extreme nutrient values at the beginning of the investigating period indicated a possible nutrient flux. Nutrient and temperature were recognized as the main environmental drivers of phytoplankton and bacterioplankton community abundance.

Bacterial transcriptional response to labile exometabolites from photosynthetic picoeukaryote Micromonas commoda

Dissolved primary production released into seawater by marine phytoplankton is a major source of carbon fueling heterotrophic bacterial production in the ocean. The composition of the organic compounds released by healthy phytoplankton is poorly known and difficult to assess with existing chemical methods. Here, expression of transporter and catabolic genes by three model marine bacteria (Ruegeria pomeroyi DSS-3, Stenotrophomonas sp. SKA14, and Polaribacter dokdonensis MED152) was used as a biological sensor of metabolites released from the picoeukaryote Micromonas commoda RCC299. Bacterial expression responses indicated that the three species together recognized 38 picoeukaryote metabolites. This was consistent with the Micromonas expression of genes for starch metabolism and synthesis of peptidoglycan-like intermediates. A comparison of the hypothesized Micromonas exometabolite pool with that of the diatom Thalassiosira pseudonana CCMP1335, analyzed previously with the same biological sensor method, indicated that both phytoplankton released organic acids, nucleosides, and amino acids, but differed in polysaccharide and organic nitrogen release. Future ocean conditions are expected to favor picoeukaryotic phytoplankton over larger-celled microphytoplankton. Results from this study suggest that such a shift could alter the substrate pool available to heterotrophic bacterioplankton.

Photosynthetic picoeukaryotes significantly affect global oceanic meta-metabolism: cellular and genome streamlining confer ecological success

Photosynthetic picoeukaryotes (PPEs) are characterized by a reduction in cell and genome size but are free living, in contrast to many other organisms that have undergone such reductions. The relative abundance of PPEs in the oceans remains to be determined, as do the evolutionary imperatives behind their cell and genome reduction. Their enigmatic nature may be deciphered through metagenomics approaches; consequently, we utilized shotgun data from the Tara Oceans database to better understand both their ecological and genomic features. The clustering of meta-metabolomic networks constructed from shotgun data from 10 different sampling sites was influenced by the proportion of PPEs in the data sets. This, along with the relative abundance of RUBISCO sequences belonging to PPEs, indicates that they have a significant effect on oceanic meta-metabolism, emphasizing the evolutionary success of the streamlining strategy. Using rRNA sequences extracted from the shotgun data, a global oceanic distribution of PPEs showed little variation, including those lineages with reduced genome sizes. This indicates that genome and cellular streamlining is not an adaptation to environmental parameters but may rather be a community-driven effect. Lastly, and surprisingly given their role as primary producers, PPEs were found to comprise only 2 to 49% (17% on average) of all picoeukaryotes across 93 metagenomes. We show that contamination of the data set by eukaryotes with larger cell sizes is not responsible for the anomaly, and so the observation remains to be explained. The approaches described here allow us to draw a direct link between taxonomic composition and meta-metabolomic capacity, with implications for better understanding carbon fixation, biogeochemical cycling, and planetary self-regulation.

Distinct bacterial communities associated with photosynthetic picoeukaryotes in Lake Dongting throughout the seasonal cycle as revealed by flow cytometry sorting

Associations and interactions between bacteria and phytoplankton are prevalent in aquatic systems. However, the coexistence of bacteria with the smallest phytoplankton groups is mostly unknown. Here, the associated bacteria (AB) of photosynthetic picoeukaryotes (PPEs) in the Yangtze-connected Lake Dongting were studied using flow cytometry sorting combined with high-throughput sequencing of the 16S rRNA gene throughout the seasonal cycle. Bacteria in the corresponding whole water (WWB) were also investigated for comparison. Our results revealed a distinct bacterial community of AB compared to WWB. The taxonomic and phylogenetic diversity indices, as well as the richness, of AB were significantly lower than those of WWB; however, AB showed higher Pielou’s evenness. Proteobacteria (53.8%) was the most abundant phylum in AB, whereas Actinobacteria was predominant in WWB (36.0%). Several abundant clades, including Flavobacterium, Pseudomonas, Rheinheimera, Novosphingobium, and Sphingomonas, were significantly enriched in AB. Functional prediction analysis identified 116 KEGG pathways differing significantly between AB and WWB. Particularly, functional properties involved in cellular processes (mainly bacterial chemotaxis), environmental information processing, organismal systems and xenobiotics biodegradation and metabolism were greatly enriched in AB. Total nitrogen, temperature and chlorophyll a correlated significantly with the community dissimilarity between AB and WWB. Our results shed new light on the bacterial diversity and function associated with the smallest phytoplankton.

Composition and genetic diversity of picoeukaryotes in the northeastern South China Sea during the Luzon winter bloom

We analyzed picoeukaryotes during the Luzon winter bloom, in combination with remote sensing, flow cytometry and high throughput sequencing. Satellite images showed a seasonal increase in phytoplankton in the study area during winter, and the in situ campaign observed an extremely high abundance of photosynthetic picoeukaryotes, which were the main contributors to the total phytoplankton biomass. Auto- and mixotrophic picoalgae (Chlorophyta, Ochrophyta, and Dinophyta), heterotrophic flagellates (Radiolaria) and parasites (Syndiniale) were found to be potentially numerous components of picoeukaryote communities. Overall, auto- and mixotrophic photosynthetic picoeukaryotes were mainly distributed in the upper layers (0–100 m) while heterotrophic flagellates and parasites dominated the deep layers (200–300 m). The Kuroshio-induced upwelling stimulated the growth of photosynthetic picoeukaryotes, which mainly belonged to Chlorophyta, especially the class Mamiellophyceae.

The Spatial Variation in Chlorophyte Community Composition From Coastal to Offshore Waters in a Subtropical Continental Shelf System

Globally, the continental shelf occupies less than 10% of the total sea surface, but supports substantial primary production and fisheries. Photosynthetic picoeukaryotes (PPE) are important primary producers in marine ecosystems, and chlorophytes make a significant contribution to PPE abundance. Although the distribution of chlorophytes has been widely studied, little is known about how their community composition varies along the coastal-offshore gradient in subtropical continental shelf waters. To better understand their spatial variations, we employed metabarcoding data of 18S rRNA V4 gene to examine chlorophyte composition within the surface and deep chlorophyll maximum (DCM) layers over the continental shelf in the East China Sea (ECS) with high-resolution sampling. Our results indicate a higher chlorophyte diversity in the surface layer than in the DCM. In addition, we found that chlorophytes in oligotrophic Kuroshio surface water were the most diverse, with a higher percentage of Chloropicophyceae, prasinophyte clade V, prasinophyte clade IX, Palmophyllophyceae and Chlorodendrophyceae. In more eutrophic waters along the coast and within the DCM layer, chlorophytes were dominated by Mamiellophyceae. A significant spatial variation in community composition occurred along the nutrient gradient in the subtropical continental shelf ecosystem, varying from Mamiellophyceae-dominant in coastal water to Chloropicophyceae-dominant in offshore water. Furthermore, at a low saline coastal station, which had the lowest chlorophyte diversity, Picochlorum (Trebouxiophyceae) was dominant. Overall, chlorophyte diversity was positively correlated with temperature, and negatively correlated with silicate concentration. This study revealed that terrestrial input into continental systems influences chlorophyte diversity in the subtropical northwestern Pacific Ocean.

Differences in the Composition of Abundant Marine Picoeukaryotes in the Marginal Sea Derived from Flooding

The transient impact of flooding on the community composition of marine picoeukaryotes (PEs, cell size ≤5 μm) in the East China Sea (ECS) was revealed in this study. In a summer without flooding (i.e., July 2009), photosynthetic picoeukaryotes (PPEs) were more abundant in the area covered by the Changjiang River diluted water (CDW, salinity ≤31) than in the non-CDW affected area. According to the 18S ribosomal RNA phylogeny, Alveolata (all from the superclass Dinoflagellata) was the main community component accounting for 72 to 99% of the community at each sampling station during the nonflooded summer. In addition to Dinoflagellata, diatoms or Chlorophyta also contributed a considerable proportion to the PE assemblage at the stations close to the edge of CDW coverage. In July 2010, an extreme flooding event occurred in the Changjiang River basin and led to the CDW covering nearly half of the ECS. In the flooded summer, the abundance of PPEs in the CDW-covered area decreased significantly to less than 1 × 104 cells ml-1. Compared to that during the nonflooded summer, the diversity of the PE composition was increased. While Dinophyceae still dominated the surface waters, Syndiniophyceae, which were represented by the uncultured Marine Alveolata Group (MALV)-I and MALV-II, accounted for a substantial amount in the Dinoflagellata superclass relative to this community composition in the nonflooded summer. Furthermore, a variety of plankton, including Cryptophyta, Haptophyta, Picobiliphyta, the uncultured Marine Stramenopiles (MASTs) and heterotrophic nanoflagellates, were observed. The nutrition modes of these PEs have been reported to be mixotrophic or heterotrophic. Therefore, it was inferred that the potentially mixotrophic and heterotrophic PE compositions might be favored in the marginal sea in the flooded summer.

Optimization of Preservation Methods Provides Insights into Photosynthetic Picoeukaryotes in Lakes.

ABSTRACTAs the key contributor to plankton biomass and nutrient cycling in aquatic ecosystems, photosynthetic picoeukaryotes (PPEs) have been recently investigated in freshwater ecosystems. However, the limited access to remote areas creates challenges for PPE sample preservation before sorting and counting by flow cytometry (FCM) in the laboratory. Here, we explored the effects of different preservation methods on the PPE community by combining FCM sorting and high-throughput sequencing. Our results showed that dimethyl sulfoxide (DMSO) cryoprotection could destroy the fluorescence and cell structure of the PPEs, making the subsequent FCM analysis and sorting difficult. Aldehyde fixation maintained the PPE fluorescence, and the fixed samples were of sufficient quality for abundance analysis and sorting by FCM. However, the sequencing results showed that, after preservation by aldehydes, the proportion of PPEs dramatically decreased to approximately 10%, in comparison to 90% in the fresh samples, and the sequences of Ascomycota significantly increased. In contrast, preservation with Pluronic F68 (F68) not only could maintain the PPE abundance close to the initial value but also could keep the PPE community similar to that in the fresh samples over a storage time of 6 months. Thus, F68 cryopreservation is a suitable preservation method for PPE communities from freshwater lakes.IMPORTANCE PPEs contribute significantly to primary productivity in freshwater ecosystems. The combination of FCM sorting and high-throughput sequencing has been shown to be a powerful approach and can largely improve our view of the PPE diversity. However, the water samples could not be counted and sorted immediately after sampling from many lakes due to the inaccessibility of FCM in the field. Thus, the comparison of different preservation methods that allow subsequent analysis of the community structure by high-throughput sequencing is an urgent need. Our results indicated that F68 cryopreservation could maintain the PPE abundance close to the initial value and keep the community similar to that in the fresh samples over a storage time of 6 months.

Seasonal and Spatial Variations in Synechococcus Abundance and Diversity Throughout the Gullmar Fjord, Swedish Skagerrak.

The picophytoplankton Synechococcus is a globally abundant autotroph that contributes significantly to primary production in the oceans and coastal areas. These cyanobacteria constitute a diverse genus of organisms that have developed independent niche spaces throughout aquatic environments. Here, we use the 16S V3–V4 rRNA gene region and flow cytometry to explore the diversity of Synechococcus within the picophytoplankton community in the Gullmar Fjord, on the west coast of Sweden. We conducted a station-based 1-year time series and two transect studies of the fjord. Our analysis revealed that within the large number of Synechococcus amplicon sequence variants (ASVs; 239 in total), prevalent ASVs phylogenetically clustered with clade representatives in both marine subcluster 5.1 and 5.2. The near-surface composition of ASVs shifted from spring to summer, when a 5.1 subcluster dominated community developed along with elevated Synechococcus abundances up to 9.3 × 104 cells ml–1. This seasonal dominance by subcluster 5.1 was observed over the length of the fjord (25 km), where shifts in community composition were associated with increasing depth. Unexpectedly, the community shift was not associated with changes in salinity. Synechococcus abundance dynamics also differed from that of the photosynthetic picoeukaryote community. These results highlight how seasonal variations in environmental conditions influence the dynamics of Synechococcus clades in a high latitude threshold fjord.

Significant Effects of Associated Microorganisms on the Community of Photosynthetic Picoeukaryotes.

Photosynthetic picoeukaryotes (PPEs) form associations with other microorganisms. However, whether and how the associated microbes affect PPE communities remain unknown. We used flow cytometric cell sorting combined with parallel high-throughput sequencing of the 18S and 16S rRNA genes to simultaneously investigate PPEs and their associated microbial communities in the Yangtze-connected Lake Dongting. The lake harbors a great diversity of PPEs. PPE communities exhibited significant temporal rather than spatial variations. Two distinct PPE taxa affiliated with Discostella nipponica and Poterioochromonas malhamensis were dominant during winter/spring and summer, respectively. Parallel high-throughput sequencing revealed a great diversity of associated bacteria and non-pigmented eukaryotes (NPEs) in PPEs sorts. Proteobacteria, Actinobacteria, Bacteroidetes, and Cyanobacteria among the associated bacteria and fungi among the associated NPEs were dominant. PPEs were more apparently associated with bacteria than with NPEs. The co-occurrence network of PPEs and associated microbes formed five major modules, which exhibited distinct temporal patterns, being specific to a certain period. Variations in PPEs communities were significantly correlated with both environmental factors and associated microbial communities. In variation partitioning analysis, the associated bacteria explained the greatest variations in PPE communities, and associated bacteria and NPEs co-explained a large portion of environmental effects on PPE communities. Our results highlight the significance of associated microbes in shaping PPE communities.

Seasonal Dynamics of Photosynthetic Picoeukaryotes in a Monsoon-Influenced Tropical Bay: a Flow Cytometric and Chemotaxonomic Approach.

The composition and ecology of photosynthetic picoeukaryotes (PPE) are essential for understanding microbial food web functioning. We hypothesize that the simultaneous use of flow cytometry (FCM) and high-performance liquid chromatography (HPLC) tools will aid in discerning the dominant PPE groups contributing to abundance and biomass under prevailing environmental conditions. The PPE seasonal community abundance and pigment biomass were investigated from a southwest monsoon-influenced tropical bay from June 2015 to May 2016. A size-fractionated (<3 µm) approach using FCM and HPLC revealed five and six PPE groups, respectively. Picocryptophytes dominated the PPE biomass under varied environmental conditions, whereas picodiatoms contributed substantially, being abundant under turbulent, low-temperature, nutrient (NO3-, SiO44-)-enriched conditions. The picochlorophytes dominated the community numerically. The relatively higher abundance and biomass of picoprasinophytes and a positive correlation with NO3- and NH4+ imply proliferation under higher nutrient concentrations. The least contributors to biomass were dinoflagellates and picoprymnesiophytes. The relatively larger cell size of picocryptophytes and picodiatoms resulted in higher cumulative biomass, signifying their role in the microbial food web. Our study proposes incorporation of additional indicator pigments in algorithms used to estimate coastal picophytoplankton contribution to total phytoplankton biomass to avoid discrepancies.

Elemental and macromolecular composition of the marine Chloropicophyceae, a major group of oceanic photosynthetic picoeukaryotes

AbstractChloropicophyceae (Prasinophyte Clade VII) are small nonmotile coccoid cells with cell diameters ranging from 1 to 3 μm. Molecular surveys indicate they are relatively high in abundance in moderately oligotrophic oceanic waters and may substantively contribute to biogeochemical cycling in the sea. Here, we quantify the elemental and macromolecular composition of three subtropical Chloropicophyceae strains: Chloropicon mariensis, Chloropicon maureeniae, and Chloropicon roscoffensis under nutrient‐sufficient exponential growth and nitrate starvation. Under nutrient‐sufficient conditions the Chloropicophyceae are high in C : N and quite low in C : P and N : P relative to the canonical Redfield ratio, reflecting their relatively high nucleic acid composition compared to many other phytoplankton taxa. Nitrate starvation causes increases in C : N and C : P and decreases in N : P, primarily due to increases in carbohydrate and lipid and decreases in protein and RNA. There is genetic evidence that unlike most other green algae, Chloropicophyceae are diploid. The high nucleic acid content in the Chloropicon is consistent with the hypothesis that the nucleus, as a nonscalable component, takes up a larger and substantial proportion of cell mass in diploid picoeukaryotes. The elemental and macromolecular composition of these Chloropicophyceae, and relatively homeostatic response to N‐starvation compared to diatoms, provides some insight into their success in the moderately oligotrophic ocean.

Cryptophyte and Photosynthetic Picoeukaryote Abundances in the Bransfield Strait during Austral Summer

A remarkable shift in the species composition and size distribution of the phytoplankton community have been observed in coastal waters along the Antarctic Peninsula over the last three decades. Smaller photoautotrophs such as cryptophytes are becoming more abundant and important for the regional ecosystems. In this study, flow cytometry was used to quantify the smallest phytoplankton in the central Bransfield Strait and explore their distribution across the strait in relation to physical and chemical properties of the two major water masses: the warmer and less saline Transitional Zonal Water with Bellingshausen Sea influence (TBW), and the cold and salty Transitional Zonal Water with Weddell Sea influence (TWW). Pico- and nano-phytoplankton clusters were distinguished and enumerated in the cytograms: photosynthetic picoeukaryotes, cryptophytes (about 9 µm in size), and smaller (3 µm) nanophytoplankton. It was shown that nanophytoplankton developed higher abundances and biomasses in the warmer and less saline TBW. This biotope was characterized by a more diverse community with a pronounced dominance of Cryptophyta in terms of biomass. The results support the hypothesis that increasing melt-water input can potentially support spatial and temporal extent of cryptophytes. The replacement of large diatoms with small cryptophytes leads to a significant shift in trophic processes in favor of the consumers such as salps, which able to graze on smaller prey.

Seasonality of Coastal Picophytoplankton Growth, Nutrient Limitation, and Biomass Contribution.

Picophytoplankton in the Baltic Sea includes the simplest unicellular cyanoprokaryotes (Synechococcus/Cyanobium) and photosynthetic picoeukaryotes (PPE). Picophytoplankton are thought to be a key component of the phytoplankton community, but their seasonal dynamics and relationships with nutrients and temperature are largely unknown. We monitored pico- and larger phytoplankton at a coastal site in Kalmar Sound (K-Station) weekly during 2018. Among the cyanoprokaryotes, phycoerythrin-rich picocyanobacteria (PE-rich) dominated in spring and summer while phycocyanin-rich picocyanobacteria (PC-rich) dominated during autumn. PE-rich and PC-rich abundances peaked during summer (1.1 × 105 and 2.0 × 105 cells mL–1) while PPE reached highest abundances in spring (1.1 × 105 cells mL–1). PPE was the main contributor to the total phytoplankton biomass (up to 73%). To assess nutrient limitation, bioassays with combinations of nitrogen (NO3 or NH4) and phosphorus additions were performed. PE-rich and PC-rich growth was mainly limited by nitrogen, with a preference for NH4 at >15°C. The three groups had distinct seasonal dynamics and different temperature ranges: 10°C and 17–19°C for PE-rich, 13–16°C for PC-rich and 11–15°C for PPE. We conclude that picophytoplankton contribute significantly to the carbon cycle in the coastal Baltic Sea and underscore the importance of investigating populations to assess the consequences of the combination of high temperature and NH4 in a future climate.

Evidence for mixotrophy in pico-chlorophytes from a new Picochlorum (Trebouxiophyceae) strain.

Mixotrophs are increasingly recognized for their wide distribution in aquatic ecosystems and significant contributions to biogeochemical cycling. Many taxa within the phyla Chrysophyta, Cryptophyta, and Haptophyta are capable of phago-mixotrophy, however, phagotrophy in the Chlorophyta remains controversial due to insufficient research and solid evidence. In this study, we identified a new strain, Picochlorum sp. GLMF1 (Trebouxiophyceae), using 18S rRNA gene analysis and morphological observations. It displayed multi-cell division through autosporulation (two- or four-cell daughters) and has two unequal flagella that have never been reported in the genus Picochlorum. By using multiple methods, including 3D bioimaging analysis, acidic food vacuole-like compartment staining, and prey reduction calculation, we discovered and confirmed bacterivory in Picochlorum, which provided strong evidence for phago-mixotrophy in this green alga. In addition, we found that Picochlorum sp. GLMF1 cannot grow under complete darkness or prey-depleted conditions, suggesting that both light and bacteria are indispensable for this strain, and its mixotrophic nutrition mode is obligate. Like other phago-phototrophs, Picochlorum sp. GLMF1 is capable of regulating their growth and ingestion rates according to light intensity and inorganic nutrient concentration. The confirmation of mixotrophy in this Picochlorum strain advances our understanding of the trophic roles of green algae, as well as the photosynthetic picoeukaryotes, in marine microbial food webs.